Abstract A fundamental goal of surgery is function preservation and minimizing patient morbidity. Current nerve identification during surgery utilizes non-quantifiable criteria such as anatomy, texture, color, relationship to surrounding structures to distinguish nerves from non-nerve tissues. In instances of trauma, tumor invasion or infection, nerve identification using the above criteria may be challenging. Using white light reflectance, which is the standard mode of illumination in operating rooms, the visual difference between small nerves, such as cavernosal nerves important during radical prostatectomy or distal branches of the facial nerve important during surgery for salivary gland neoplasms, and adjacent tissue can be imperceptible. Our laboratory has extensive experience in developing molecularly targeted smart probes for surgical visualization of nerves. Recently, we have identified a novel peptide (HNP401) via phage display to have high level of human nerve binding. In this proposal, we aim to optimize HNP401 to enable its clinical translation for human surgery.